Abstract
Objective:
To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain.
Methods:
According to the published SARS coronavirus genome sequences, the full length cDNA of N protein from SARS coronavirus PUMC2 strain was cloned by RT-PCR and the cDNA was cloned into the pET32a expression vector. The recombinant N protein was expressed in E. coli BL21 (DE3), and purified by Ni(2+)-NTA.
Results:
Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained.
Conclusions:
The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Cloning, Molecular*
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Coronavirus Nucleocapsid Proteins
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DNA, Complementary / genetics
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DNA, Viral / genetics*
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Escherichia coli / genetics
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Genetic Vectors
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Genome, Viral
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Molecular Sequence Data
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Nucleocapsid Proteins / biosynthesis
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Nucleocapsid Proteins / genetics*
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Nucleocapsid Proteins / isolation & purification
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RNA, Viral / genetics
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Sequence Analysis, DNA
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Severe acute respiratory syndrome-related coronavirus / genetics*
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Severe acute respiratory syndrome-related coronavirus / isolation & purification
Substances
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Coronavirus Nucleocapsid Proteins
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DNA, Complementary
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DNA, Viral
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Nucleocapsid Proteins
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RNA, Viral
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Recombinant Fusion Proteins